Department of interventional radiology, The Affiliated Cancer Hospital of Guizhou Medical University, Guiyang 550002, PR China.
Department of Anesthesiology, Guizhou Medical University, Guiyang 550002, PR China.
Res Vet Sci. 2021 May;136:377-384. doi: 10.1016/j.rvsc.2021.03.003. Epub 2021 Mar 3.
Osteoarthritis is currently one of the most common chronic diseases. As life expectancy increases, its prevalence and incidence are expected to rise. At present, more and more evidences prove the correlation between the complement system and osteoarthritis (OA). This study aims to investigate complement C5's influence on the effect of MK801 on osteoarthritis synovial fibroblasts (OA-SFs).
We used IL-1b to induce OA-SFs derived from mice to obtain OA-SFs. And we performed RT-PCR and Western Blot assays to evaluate the expression levels of associated mRNA and protein. The alteration of MAC expression on OA-SFs cell membrane was evaluated by immunofluorescence assay. The expression of related inflammatory factors of OA-SFs was evaluated by ELISA experiment.
MK801 could significantly inhibit the expression of osteoarthritis (OA) marker factors, such as: membrane attack complex (MAC), tumor necrosis factor-α (TNF-α) and matrix metalloproteinase-13 (MMP13). Meanwhile, MK801 can significantly inhibit the expression of complement C5 (C5) in OA-SFs. Immunofluorescence assay showed that MAC expression on OA-SFs cell membrane was significantly inhibited by MK801. The nucleo-plasmic separation experiment demonstrated that MK801 could significantly inhibit the activation of Nuclear factor-κB (NF-κB) signaling pathway in OA-SFs. Futhermore, koncking down the expression of C5 reversed the inhibition MK801 on the expression of OA-SFs inflammatory factors.
These results illustrated two points: first, MK801 inhibited the generation of MAC and the release of inflammation factors in OA-SFs through C5; second: MK801 inhibited the activation of NF-κB signaling pathway in OA-SFs.
骨关节炎目前是最常见的慢性疾病之一。随着预期寿命的增加，其患病率和发病率预计将上升。目前，越来越多的证据证明补体系统与骨关节炎（OA）之间存在相关性。本研究旨在探讨补体 C5 对 MK801 对骨关节炎滑膜成纤维细胞（OA-SFs）作用的影响。
我们使用 IL-1b 诱导从小鼠获得的 OA-SFs 来获得 OA-SFs。并通过 RT-PCR 和 Western Blot 分析评估相关 mRNA 和蛋白的表达水平。通过免疫荧光分析评估 OA-SFs 细胞膜上 MAC 表达的变化。通过 ELISA 实验评估 OA-SFs 相关炎症因子的表达。
MK801 可显著抑制 OA 标志物因子的表达，如膜攻击复合物（MAC）、肿瘤坏死因子-α（TNF-α）和基质金属蛋白酶-13（MMP13）。同时，MK801 可显著抑制 OA-SFs 中的补体 C5（C5）表达。免疫荧光分析表明，MK801 显著抑制了 OA-SFs 细胞膜上 MAC 的表达。核质分离实验表明，MK801 可显著抑制 OA-SFs 中核因子-κB（NF-κB）信号通路的激活。此外，敲低 C5 的表达逆转了 MK801 对 OA-SFs 炎症因子表达的抑制作用。
这些结果说明了两点：首先，MK801 通过 C5 抑制了 MAC 的产生和 OA-SFs 中炎症因子的释放；其次，MK801 抑制了 OA-SFs 中 NF-κB 信号通路的激活。
